LT2 Rule
Cryp tosporidium
&
E. coll
Sample Collection
Recommendations
Pocket Crui
(Printer Friendly Version)
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Cryptosporidium
Preparation for Sample Filtration
Thoroughly clean* all reused collection equipment and containers (new tubing,
fittings, etc. should not need cleaning), and wear gloves whenever needed to
prevent sample contamination throughout the collection and filtration procedure.
Before connecting the sampling unit to the tap, thoroughly flush stagnant water
and debris in the sample line until the temperature and turbidity stabilize
(approx. 2-3 mins).
Complete the sample collection form including utility and
sample ID information, date, turbidity, pH and temperature.
Pressurized source
1. Determine sample line water pressure at the location described to the State
2. Connect assembled sampling unit, without capsule/foam filter, to sample tap
3. Flush the sampling unit for 2-3 minutes and test for leaks
Unpressurized source
1. Fill 30+ L carboy from a pressurized sample tap or manually collect the sample
using hose, pump, funnel, etc. from the location described to the State
2. Chill the full carboy to between 1° and 10°C if filtration is not performed
immediately after filling the carboy
3. Assemble sampling unit, without capsule/foam filter, and insert influent tube
into the center of the carboy
4. Turn on pump to flush the sampling unit for 2-3 minutes and test for leaks
* Refer to the cleaning procedure located on the Packing and Shipping panel
of this pocket guide
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Sample Collection Recommendafu
Cryptosporidium
Pressurized Sample Filtration
Enviroehek™ or Envirochek™ HV Capsules
1. Install filter capsule
2, Record start time and initial meter reading
3. Slowly open sample tap, bleed out air, and adjust up to
the following maximum values:
• 100 psig and 2 L/min
4. When at least 10 L has passed through the filter, turn off sample tap
allowing pressure to decrease until water stops
5. Record stop time and final meter reading
6. Hold inlet pointing up, remove outlet tubing and allow water to drain
7. Open bleed valve to speed the draining process and disconnect inlet tubing
8. Seal the capsule with vinyl end caps, close the bleed valve, and place in
gallon-size zippered plastic bag
9, Chill the sample as soon as possible to between 1° and 10°C
until ready for shipment
Tap pressure >1QO psig
Pressure
regulator
Envirochek™ Flow totalizer T
capsule
Tap pressure 1-100 psig
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Filta-Max© Filter
1 . Install foam filter inside filter housing
2. Record start time and initial meter reading
3. Slowly open sample tap and adjust up to the following recommended values:
• 72.5 psig and 3-4 Umin
4. When at least 1 0 L has passed through the filter, turn off sample tap allowing
pressure to decrease until water stops
5. Record stop time and final meter reading
6. Hold inlet pointing up, remove the outlet tubing and allow water to drain
7. Disconnect the intet tubing
8 Seal filter inside filter housing with rubber stoppers and place into a
gallon-size zippered plastic bag
9, Chill the sample as soon as possible to between 1° and 10°C
until ready for shipment
Tap pressure >72,5 psig
Pressure
regulator
Filla-Ma*®
filter housing
Tap pressure 7.5 - 72,5 psig
Fife-Max® filter housing
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Cryptosporidium
Unpressurized Sample Filtration
Envirochek™ or Envirochek™ HV Capsules
1. Install filter capsule and turn on pump
2, Record start time and initial meter reading
3. Bleed out air and adjust flow rate up to a maximum of 2 L/min
4. When at least 10 L has passed through the filter, turn off pump
5. Record stop time and final meter reading
6. Hold inlet pointing up, remove outlet tubing and allow water to drain
7. Open bleed valve to speed the draining process and disconnect inlet tubing
8. Seal the capsule with vinyl end caps, close the bleed valve, and place in
gallon-size zippered plastic bag
9, Chill the sample as soon as possible to between 1° and 10°C
until ready for shipment
Filta-Max® Filter
PerisEallic pump
I
Flow rate meter
with valve x i
1. Install foam filter inside filter housing and
turn on pump
2. Record start time and initial meter reading
3. Adjust flow rate up to a maximum of 3-4 L/min
4. When at least 10 L has passed through the filter, turn off pump
5. Record stop time and final meter reading
6. Hold inlet pointing up, remove outlet tubing and allow water to drain
7. Disconnect inlet tubing
8. Seal filter inside filter housing with rubber stoppers and place into a
gallon-size zippered plastic bag
9. Chili the sample as soon as possible to between 1° and 10°C until ready
for shipment
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Cryp tosporidium
10-L Bulk Water Collection
Thoroughly clean* all reused collection equipment and containers (new
hoses, cubitainers, etc, should not need cleaning), and wear gloves
whenever needed to prevent sample contamination throughout the
collection procedure. Prior to sample collection, thoroughly flush stagnant
water and debris in the sample line until the temperature and turbidity
stabilize (approx. 2-3 mins).
Complete the sample collection form including utility and sample
ID information, date, time, turbidity, pH, and temperature.
j
1. Fill the 10-L cubitainer from a pressurized
sample tap or manually collect sample using
hose, pump, funnel, etc. from the location
described to the State.
2. Once the sample is collected,
immediately place the cap on the
cubitainer and tighten
3. Chill the sample as soon as possible to
between 1" and 10°C before shipment
Refer to the cleaning procedure located on the Packing and
Shipping panel of this pocket guide
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Sample Collection Recommendatu
Cryptosporidium
Matrix Spike Sample Collection
Send an extra 10 L bulk sample for spiking at the Cryptosporidium laboratory
concurrent with the 1st and 21st routine monitoring sample collections and
again with every 20 samples if more than 40 source water samples are
collected. Coordinate the collection of matrix spike samples with your laboratory.
Matrix spike samples should be collected from the same location and at the
same time as the associated routine sample. The matrix spike sample and
routine sample should be the same volume. Label the cubitainer with the
following information using a waterproof pen: PWSID, facility name, and sample
collection date. Chill the sample as soon as possible to between 1° and 10°C
before shipment.
10-L Matrix Spike Sample
Follow instructions for collecting a
10-L bulk water sample given on
previous panel.
Matrix Spike Samples Greater
than 10-L
Either ship the entire matrix spike volume
as a bulk sample to the laboratory to be spiked and analyzed, OR filter all
but 10 L of the matrix spike sample at your utility and collect the remaining
10 L as a bulk sample. Chill both the filter and bulk sample as soon as possible
to between 1° and 10°C before shipment. Clearly label both the filter and
bulk sample as comprising 2 parts of a single sample.
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9ns:
ocket Guide
E. coli
Sample Collection
Wear gloves whenever needed to prevent
sample contamination throughout the
collection procedure.
Complete the sample collection form
including utility and sample ID
information, date and time, etc.
1. Water taps used for sampling
should be free of aerators, hose
attachments, etc.
2. Prior to sample collection, thoroughly
flush stagnant water and debris
in the sample line until the
temperature and turbidity
stabilize (approx, 2-3 mins):
DO NOT rinse the sample
bottle
3. Record the turbidity,
pH, and temperature
4. Aseptically fill the sterile
E. coli sample bottle
from a pressurized
sample tap or manually
collect sample using,
hose, pump, funnel,
etc, from the location
described to the State
5. Leave at least one inch of
head space in the E. coli sample
bottle, if possible. Collect at least 100 ml of sample for analysis
6. Immediately close the sample bottle
7. Chill sample as soon as possible to between 0" and 10'C until ready
for immediate shipment
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Packing and Shipping
1. Remove sample (filter, cubitainer, or sample bottle) from refrigerator or
cooler just prior to shipping. Wrap sample in bubble wrap to prevent freezing
2. Place sample in cooler lined with two large plastic trash bags
3. Pack cooler with sealed ice bags or the equivalent number of gel ice packs
sufficient to maintain the sample between 1° and 10°C during shipment. Use
the amount of coolant appropriate for the season, geographic location, and
cooler-type used (follow laboratory recommendations). NEVER USE DRY ICE.
4. Knot and seal the two large plastic trash bags
5. Sign and date sample collection form, place in a water-tight bag in cooler
6. Seal and secure cooler with tape
7. Complete airbill and attach to cooler, retain shipper's copy
8. Ship sample(s) to the laboratory via overnight delivery
^Recommended Sampling Equipment Cleaning Procedure
(or alternatively follow the manufacturer's cleaning instructions)
Not for sterilization of E. colt sample bottles
• Thoroughly clean all reused influent equipment prior to the capsule/filter
housing/bulk water container with a warm detergent solution and scrubbing
with a brush (new equipment should not need cleaning)
• Expose equipment to hypochlorite solution (Add 25 mL of 5% household
bleach for every 1 gallon of pH 7 water) for at least 30 minutes at
room temperature
• Thoroughly rinse equipment with reagent grade water that is free of analytes
and interfering substances. Dry equipment in an area free of potential
Cryptosporidium contamination
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This pocket guide Is intended as an outline and supplement to the
Source Water Monitoring Guidance Manual for Public Water Systems
and the on-line Sample Collection Training Module. This guide is
intended to provide useful information and recommendations, but
does not substitute for the LT2 Rule.
The guidance manual is available for download and study at:
http://www.epa,gov/safewater/disinfection/lt2/pdfs/guide_lt2_
swmonitoringguidance.pdf
The on-line training module can be accessed at:
http://www.epa.gov/safewater/lt2/training/modules.html
Copies of this pocket guide can be ordered by using the document
number listed below and contacting the National Service Center
for Environmental Publications at 1-800-490-9198
Office of Water (46Q1M)
EPA 817-K-06-002
April 2006
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